Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more clearly understood, the present invention will be described in further detail with reference to the accompanying drawings and the following detailed description.
The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those skilled in the art without creative efforts belong to the protection scope of the present invention.
In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience of description and to simplify the description, but do not indicate or imply that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore should not be construed as limiting the present invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically limited otherwise.
In the present invention, unless otherwise expressly specified or limited, the terms "mounted," "connected," and "secured" are to be construed broadly and can, for example, be connected or detachably connected or integrated; can be mechanically or electrically connected; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meaning of the above terms in the present invention can be understood according to specific situations by those skilled in the art.
In the present disclosure, unless expressly stated or limited otherwise, the first feature "on" or "under" the second feature may comprise direct contact between the first and second features, or may comprise contact between the first and second features not directly. Also, the first feature being "on," "above" and "over" the second feature includes the first feature being directly on and obliquely above the second feature, or merely indicating that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature includes the first feature being directly under and obliquely below the second feature, or simply meaning that the first feature is at a lesser elevation than the second feature.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above should not be understood to necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples described in this specification can be combined and combined by one skilled in the art.
As shown in fig. 1-5, the cell transfer observing mechanism provided in this embodiment can be applied to cell culture and preparation processes to realize automatic transfer of the culture bottle, so that the cells in the culture bottle are placed under a microscope, and the position of the culture bottle can be automatically changed to perform comprehensive observation of the cells, so as to realize automatic movement of the culture bottle and comprehensive observation of the growth of the cells, improve the observation efficiency, avoid contact with people in the whole moving and observing processes, further reduce the risk of contamination, and reduce the cost.
Referring to fig. 1, the cell transfer observing mechanism includes a frame (not shown), a transferring assembly and an observing assembly, wherein the transferring assembly includes a suction cup assembly and a multi-directional moving assembly, the suction cup assembly is connected with the multi-directional moving assembly, and the multi-directional moving assembly is connected with the frame. The observation assembly comprises a microscope assembly and a culture bottle moving assembly, the culture bottle moving assembly comprises a moving structure and a bearing structure, the bearing structure is connected to the moving structure, the moving structure is connected to the support, and the microscope assembly is connected to the support.
When the cell culture result is observed to needs, behind the sucking disc subassembly absorption blake bottle 60, multidirectional removal subassembly work is in order to remove sucking disc subassembly and blake bottle 60 for blake bottle 60 removes to bearing structure on, and the sucking disc subassembly loosens the actuation to blake bottle 60, and bearing structure moves under the drive that removes the structure, so that the cell of microscope subassembly in to the blake bottle is observed comprehensively.
Sucking disc subassembly and multidirectional removal subassembly cooperation can realize taking the blake bottle and remove the blake bottle to the position at microscope place to in carrying out the detection and the observation of cell, the automatic transfer blake bottle is in order to reach the purpose of automatic transfer cell, improves whole observation efficiency.
The culture bottle 60 is placed on the bearing structure and moved back and forth by the moving structure, so that the culture bottle 60 is located at different positions, and then the microscope assembly can acquire all cell images in the culture bottle 60, so as to realize comprehensive observation.
In an embodiment, referring to fig. 2 and fig. 3, the supporting structure includes a supporting frame 250 and a stand, the supporting frame 250 is provided with a limiting through slot, the culture bottles 60 are placed in the limiting through slot, the supporting frame 250 is connected to the stand, and the stand is connected to the moving structure.
The limiting through groove can play a role in positioning and limiting the culture bottle 60. The carriage 250 may be positioned just above the microscope 210 to facilitate viewing of the cells.
An inclined plane is arranged on one side wall of the limiting through groove, and the bottle mouth of the culture bottle 60 is abutted to the inclined plane to play a role in positioning the culture bottle 60.
In one embodiment, referring to fig. 3, the upper end of the stand is connected with a connecting piece 244, and the connecting piece 244 is connected with the carriage 250.
In addition, a third connecting plate 241 is provided at the lower end of the stand, and the third connecting plate 241 is connected to the moving structure, and specifically, the third connecting plate 241 may be connected to the moving structure using a fastening member such as a bolt.
In an embodiment, the vertical frame includes a vertical rod 242 and a horizontal rod 243, the horizontal rod 243 is connected to the upper end of the vertical rod 242, the connecting plate 244 is connected to the outer end of the horizontal rod 243, the third connecting plate 241 is connected to the lower end of the vertical rod 242, and the vertical frame with a 7-shaped structure formed by the horizontal rod 243 and the vertical rod 242 is arranged, so that the culture bottle 60 does not collide with the microscope assembly in the moving process, and the success rate in the whole cell observation process can be improved.
In an embodiment, please refer to fig. 3, the moving structure includes a Y-axis moving structure and an X-axis moving structure, the Y-axis moving structure is connected to the support, and the X-axis moving structure is connected to the Y-axis moving structure; the stand is connected to the X-axis moving structure.
Utilize X axle to remove structure and Y axle and remove the automatic movement of structure realization to blake bottle 60 to can observe the cultivation condition of all cells in blake bottle 60 under microscope 210, so that detect the growth condition of cell in blake bottle 60 comprehensively, and whole removal process is carried out in a confined space, can not contact with the people, and then reduces the pollution risk.
In an embodiment, referring to fig. 2, the X-axis moving structure includes a first X-axis linear module 230, the first X-axis linear module 230 is connected to the Y-axis moving structure through a mounting plate 231, and the stand is connected to the first X-axis linear module 230.
In this embodiment, the first X-axis linear module 230 is provided with a first X-axis slider, and the third connecting plate 241 is connected to the first X-axis slider, so that the first X-axis slider moves in the X-axis direction under the driving of the first X-axis linear module 230, thereby moving the culture flask 60 in the X-axis direction.
In an embodiment, referring to fig. 2, a blocking plate 232 is disposed on one side of the mounting plate 231. The stop piece 232 can prevent the first X-axis linear module 230 from directly colliding with the edge of the Y-axis moving structure during the moving process.
In an embodiment, referring to fig. 2, the Y-axis moving structure includes a first Y-axis linear module 220, the first Y-axis linear module 220 is connected to the bracket, and the mounting plate 231 is connected to the first Y-axis linear module 220.
In this embodiment, the first Y-axis linear module 220 is provided with a first Y-axis slider, the mounting plate 231 is connected to the first Y-axis slider, and the first Y-axis slider is driven by the first Y-axis linear module 220 to move in the Y-axis direction, so as to move the culture bottle 60 in the Y-axis direction, and the culture bottle 60 can move in the Y-axis direction and the X-axis direction by cooperating with the first X-axis linear module 230.
In this embodiment, the first X-axis linear module 230 is an electric servo module, and the first Y-axis linear module 220 is an electric servo module.
In an embodiment, referring to fig. 2, the microscope assembly includes a microscope 210 and a controller 211, the microscope 210 is connected to the bracket through a microscope mounting bracket 212, a through slot is disposed at a lower end of the microscope mounting bracket 212, and one end of the Y-axis moving structure is disposed in the through slot. The through slots are arranged to allow the culture bottles 60 to move onto the microscope 210 under the driving of the first Y-axis linear module 220 and the first X-axis linear module 230. In addition, a notch is formed at one end of the microscope mounting bracket 212, so that the first Y-axis linear module 220 and the first X-axis linear module 230 can work conveniently, and the combined motion of the first Y-axis linear module 220 and the first X-axis linear module 230 can move the culture bottle 60 to any point within a certain range, so that the microscope 210 can observe and image at multiple points conveniently.
The microscope 210 can take a picture of the culture bottle 60 and send the image data of the cells in the culture bottle 60 to the controller 211 for observation and calculation of the culture result, thereby completing automatic observation of the whole cell culture result.
Since the stem cells in the culture flask 60 are colorless transparent objects, it is necessary to enlarge the stem cells by the microscope 210 arranged upside down to observe the growth form.
In one embodiment, the first X-axis linear module 230 may drive the X-axis slider to slide by connecting a lead screw to a servo motor through a coupling, the first Y-axis linear module 220 is fixed on the X-axis slider, and the first Y-axis linear module 220 also drives the Y-axis slider to slide by connecting the lead screw to the servo motor through the coupling; the first X-axis linear module 230 and the first Y-axis linear module 220 are combined to move the culture bottle 60 on the plane within the XY-axis moving range. The carrier 250 is hollow to facilitate imaging under the microscope 210.
In this embodiment, the embedded controller 211 may further receive and control the movement of the first X-axis linear module 230 and the first Y-axis linear module 220 through a communication manner, trigger the camera on the microscope 210 to take a picture, and upload the picture and the detection result to the controller 211.
In this embodiment, the microscope mounting bracket 212 is connected to a camera and an illumination lamp.
Foretell observation subassembly, through setting up bearing structure for bear blake bottle 60, drive blake bottle 60 by moving structure and move in X axle direction and Y axle direction, and invert microscope 210, alright observe comprehensively to the cell in the blake bottle 60, realize automatically move blake bottle 60, improve observation efficiency, so that detect the growth condition of cell in the blake bottle 60 comprehensively, and whole removal process is not contacted with the people, and then reduce the pollution risk.
In one embodiment, the multi-directional moving assembly includes a rotating assembly, a Z-axis linear module 110, a second X-axis linear module 120, and a second Y-axis linear module 130, wherein the Z-axis linear module 110 is connected to the bracket, the second X-axis linear module 120 is connected to the Z-axis linear module 110, and the second Y-axis linear module 130 is connected to the second X-axis linear module 120; the rotating assembly is connected with the second Y-axis linear module 130 and the chuck assembly is connected with the rotating assembly.
The cooperation of Z axle straight line module 110, second X axle straight line module 120 and second Y axle straight line module 130 can realize that the blake bottle removes in the three orientation of XYZ, and the setting of rotating assembly, can rotate to some blake bottles that need to rotate to observe the cell in the blake bottle.
In an embodiment, referring to fig. 4 and 5, the suction cup assembly includes a suction cup mounting plate 150 and a plurality of suction cups, the suction cups are respectively connected to the suction cup mounting plate 150, and the suction cup mounting plate 150 is connected to the rotating assembly.
A plurality of suckers are arranged on one sucker mounting plate 150, so that the plurality of culture bottles 60 can be sucked, and the plurality of culture bottles 60 can be sucked at one time, so that the transfer efficiency of the culture bottles 60 is improved.
The suction cup mounting plate 150 is provided with a plurality of through holes for mounting the suction cups. After the culture bottle 60 is sucked by the suction cup under negative pressure, when the culture bottle 60 is moved to a specified position, the suction cup eliminates the negative pressure, and the culture bottle 60 is placed in the bearing structure.
In one embodiment, referring to fig. 5, the rotating assembly includes a swing cylinder 140, the swing cylinder 140 is connected to a chuck mounting plate 150, and the swing cylinder 140 is connected to the second Y-axis linear module 130 through a first mounting base. The rotating structure can drive the sucker mounting plate 150 to rotate, so that the culture bottle 60 can be switched from a vertical state to a horizontal state, stacking can be better facilitated, and the practicability is high.
In an embodiment, referring to fig. 5, the first mounting base includes a first side plate 142 and a second side plate 141, the first side plate 142 is connected below the second side plate 141, the swing cylinder 140 is connected to the first side plate 142, and the second side plate 141 is connected to the second Y-axis linear module 130.
In order to make the structure of the whole mechanism more compact, the swinging cylinder 140 is arranged below the second Y-axis linear module 130, so that the culture bottle 60 can be conveniently sucked, and the culture bottle 60 has a better rotating space when rotating, and is more convenient to rotate.
In an embodiment, referring to fig. 5, the Z-axis linear module 110 is connected to the bracket through a second mounting base 111.
The Z-axis linear module 110 is an electric servo module, a Z-axis slider is disposed on the Z-axis linear module 110, and the second mounting base 111 is mounted on the bracket through a fastener such as a bolt.
In an embodiment, referring to fig. 5, the second X-axis linear module 120 is connected to the Z-axis linear module 110 through a third mounting seat 121.
In this embodiment, the second X-axis linear module 120 is an electric servo module, the third mounting seat 121 is connected to the Z-axis slider through a fastener such as a bolt, and the second X-axis linear module 120 is provided with a second X-axis slider.
In an embodiment, referring to fig. 5, the second Y-axis linear module 130 is connected to the second X-axis linear module 120 through a fourth mounting seat.
In this embodiment, the second Y-axis linear module 130 is an electric servo module, and the fourth mounting seat is connected to the second X-axis slide block by a fastener such as a bolt.
Specifically, the fourth mounting seat includes a first connecting plate 132 and a second connecting plate 131, the first connecting plate 132 is connected to the second X-axis slider, the second connecting plate 131 is connected to the first connecting plate 132, a through slot is formed in the second connecting plate 131, the motor on the second Y-axis linear module 130 is embedded in the through slot, and the second Y-axis linear module 130 is connected to the second connecting plate 131.
The suction cup assembly can be moved to a designated position by the second X-axis linear module 120, the second Y-axis linear module 130, and the Z-axis linear module 110, so as to transfer the culture bottle 60 to the carrying structure. And the assembly is rotated so that the sucker assembly and the culture bottle 60 are rotated until the culture bottle 60 is in a horizontal state, so as to meet the observation requirement of a microscope on cells in the culture bottle 60.
When the culture bottle 60 needs to be transferred to the microscope 210 for observing the growth condition of the cells, the sucker assembly moves to the position of the culture bottle 60 under the action of the multidirectional moving assembly, when the sucker contacts the culture bottle 60, the sucker works under negative pressure, after the culture bottle 60 is sucked, the multidirectional moving assembly works, the rotating assembly drives the sucker assembly and the culture bottle 60 to rotate until the culture bottle 60 rotates to the horizontal state, the multidirectional moving assembly drives the rotating assembly, the sucker assembly and the culture bottle 60 to move above the bearing structure, so that the culture bottle 60 is arranged above the bearing frame 250, the sucker assembly eliminates the negative pressure, then the sucker assembly, the moving assembly and the rotating assembly return to the initial state, the microscope assembly starts to work, so that the culture condition of all the cells in the culture bottle 60 can be observed under the microscope 210, and the growth condition of the cells in the culture bottle 60 can be comprehensively detected, and the whole moving process is executed in a sealed space and cannot be contacted with people, so that the pollution risk is reduced.
Foretell cell shifts observation mechanism, through setting up transfer unit and observation subassembly, absorb the blake bottle 60 back by transfer unit's sucking disc subassembly, drive sucking disc subassembly and blake bottle 60 by multidirectional removal subassembly and move, so that the blake bottle moves to the top of bearing frame 250, bear frame 250 under the drive of blake bottle removal subassembly, move to microscope 210 position, carry out the observation of the inside cell growth condition of blake bottle 60, realize automatically move blake bottle 60 to microscope 210 position, and can change blake bottle 60's position automatically, in order to reach the growth condition of observing the cell comprehensively, improve observation efficiency, whole removal and observation process do not contact with the people, and then reduce the pollution risk, and is with low costs.
In one embodiment, there is also provided a method of operating a cell transfer viewing mechanism, comprising:
when the cell culture result is observed to needs, behind the sucking disc subassembly absorption blake bottle 60, multidirectional removal subassembly work is in order to remove sucking disc subassembly and blake bottle 60 for blake bottle 60 removes to bearing structure on, and the sucking disc subassembly loosens the actuation to blake bottle 60, and bearing structure moves under the drive that removes the structure, so that the cell of microscope subassembly in to the blake bottle is observed comprehensively.
In addition, still include:
when the microscope assembly comprehensively observes the cells in the culture bottle 60, an image of the culture bottle 60 is obtained; performing binarization processing on the image of the culture bottle 60; extracting a background image of the cell; carrying out blackening treatment on the cell-free area in the image subjected to the binarization treatment to obtain a treated image; performing particle analysis on the processed image to obtain the percentage of each particle in the processed image area; calculating the fusion degree according to the percentage of each particle in the processed image area; and transmitting the fusion degree and the processed image.
When stem cell observation is started, the culture bottle 60 is placed on a tray of a detection mechanism, and two working modes are provided, wherein one mode is remote trigger detection, and the controller 211 can send a command through communication to start detection; the other is timing trigger detection, which can set an interval time for automatically detecting one song on the controller 211, and can automatically trigger detection to start working after the timing time is up.
The first X-axis linear module 230 and the first Y-axis linear module 220 work and drive the bearing assembly to drive the culture bottle 60 to move on the XY plane, and the rigidity of the servo motors of the first X-axis linear module 230 and the first Y-axis linear module 220 and the acceleration and deceleration time of starting and stopping are adjusted, so that the first X-axis linear module 230 and the first Y-axis linear module 220 move softly, and the vibration of the culture bottle 60 is reduced to the maximum extent. The camera and the illuminating lamp are in fixed positions and cannot shake. When the first X-axis linear module 230 and the first Y-axis linear module 220 move to the photographing position, a certain time is delayed to ensure the stationary state of the culture bottle 60 and the culture solution, the illumination lamp is turned on first, and then the camera photographs a picture. After the camera takes a picture, the first X-axis linear module 230 and the first Y-axis linear module 220 can be moved to the next working position immediately; after the camera collects the cell pictures in the culture bottle 60, the pictures are sent to the controller 211, the pictures are firstly adjusted by setting appropriate threshold values for Red, Green and Blue parameters and adjusting the threshold values of Red (Red), Green (Green) and Blue (Blue) of the pictures, so that the Red of the cells with dark yellow backgrounds can be reserved, the Green and the Blue are filtered out, and meanwhile, the background colors are unchanged, so that the binarization processing is carried out on the color images, the Red color of the cell background area is maximized, and the background of the cells is lifted; and then, carrying out blackening on the image area without the cells, carrying out particle analysis on the blacked image to obtain the percentage of each particle in the total image area, and summing to obtain the fusion degree.
After the culture flask 60 was enlarged by a 40-fold objective lens, the original image was taken by a camera, and the background light source was a halogen lamp. The cell-free area is yellow, the stem cell center is dark yellow, and the periphery is colorless. Completely blacking the background without the color of the cell-free area to ensure that the cell-free area and the cell-free area have obvious difference, and carrying out particle detection on the processed binary image without the cell to obtain the total number of particles and the area percentage of each particle in the image; the sum of the area percentages of all the particles on the image area can obtain the percentage of the cells on the surface of the culture dish in the image area, namely the fusion degree, because the photographed area is only a small area in one culture bottle 60, a plurality of images are collected on one culture bottle 60, and the most real fusion value is obtained by carrying out statistics on the results of all the image processing and averaging.
It should be noted that, as will be clear to those skilled in the art, the specific implementation process of the working method of the cell transfer observing mechanism described above can refer to the corresponding description in the previous embodiment of the cell transfer observing mechanism, and for convenience and brevity of description, no further description is provided here.
The technical content of the present invention is further described by the embodiments only, so that the reader can understand it more easily, but the embodiments of the present invention are not limited thereto, and any technical extension or re-creation according to the present invention is protected by the present invention. The protection scope of the present invention is subject to the claims.